臺灣博碩士論文加值系統

3-chloro-N-(4-methoxyphenyl)furo[2,3-b]quinolin-4-amine為一新合成具抗腫瘤活性的化合物，簡稱PK-L4，在人類癌細胞株之平均GI50為0.025 ?嵱，顯示其良好之細胞毒殺效果。靜脈給予4.7 mg/kg PK-L4溶液劑的體內分佈結果顯示PK-L4會大量累積在中樞系統，對於抗癌藥物來說，因為高濃度而造成的毒性傷害是我們不樂見的，因此藉由劑型上的開發使得抗癌藥物具有靶向作用是一大研究目標。
　　本研究中利用高效能液相層析來進行PK-L4的檢測，以C18為層析管柱，移動相則由55% 0.01M 1-pentasulfonic acid sodium salt （pH3）與45% acetonitrile組成，檢出器為電化學檢出器。
　　我們利用trimyristin、Gelucire 53/10、stearylamine、hydrogenated soybean phosphatidylcholine、polyethylene glycol monostearate （45E.Q.）製備出兩種PK-L4固體脂質微粒，顆粒大小為30 and 50 nm（含與不含 PEG 45E.Q.），且包埋率皆高達99%；透過示差熱分析儀（DSC）可以發現PK-L4與SLN的脂質有均勻的混合。體外釋放實驗顯示此兩種SLN皆為零階次的釋放，其24小時累積釋放量分別為16與30%（含與不含 PEG 45E.Q.）。體外細胞毒殺實驗發現PK-L4對於肺癌細胞具有較佳的效果，特別是經過製劑化後的固體脂質微粒處方。
　　靜脈給予PK-L4 SLN 4.7 mg/kg於健康大白鼠，並與溶液劑做比較，結果發現經過SLN製劑化的PK-L4在大部分的組織器官中，其曲線下面積與生體可用率皆較溶液劑來的好，而在動物體內的分佈尤其以肺部有明顯增加的趨勢，而於中樞的藥物累積卻明顯的降低，此結果顯示SLN的劑型對於PK-L4在動物體內的分佈上具有靶向作用的功能。

3-chloro-N-(4-methoxyphenyl)furo[2,3-b]quinolin-4-amine, PK-L4, was a novel compound with anti-tumor activity. The mean GI50 value 0.025 ?嵱 showed good cytotoxicity of PK-L4. After intravenous administration of 4.7 mg/kg PK-L4 solution to health male Wistar rats, we found that most of PK-L4 was accumulated in brain and spine.
We used trimyristin, Gelucire 53/10, stearylamine, hydrogenated soybean phosphatidylcholine and polyethylene glycol monostearate （45E.Q.）to prepare two types of PK-L4 solid lipid nanoparticle formulations. The mean diameters of two SLNs were 30 and 50 nm, with and without PEG. The entrapment efficiency of both two SLNs were approximately 99%. According to DSC result, we found that PK-L4 was homogenized in lipid. In vitro release experiment showed the PK-L4 accumulated amount of these two SLNs were 16 and 30%, with and without PEG, after 24 hours releasing. The release kinetics of two SLNs were both zero-order. In vitro cytotoxicity test showed PK-L4 had good cytotoxic effect against human lung cancer cell line, especially after formulation.
In this study, we used high performance liquid chromatography to determine the PK-L4 content in vitro and in vivo. A C18 column was used for the separation of analyte with a mobile phase consisting of 55% 0.01M pH 3 of sodium 1-pentasulfonate solution and 45% acetonitrile. We detected PK-L4 by electrochemical detector.
In most organs, compared with PK-L4 solution, the area under the curve and bioavailability of PK-L4 SLNs were all better than solution. After intravenous administration of PK-L4 SLNs, we found the accumulation of PK-L4 in lung were increased, and decreased in brain and spine. This result showed that SLN formulation might target in lung.

附圖目錄 4
附表目錄 6
中文摘要 9
Abstract 10
壹、緒論 11
一、研究背景 11
二、藥物於腫瘤的運輸 14
三、Solid lipid nanoparticles（SLN）劑型概述 15
1. SLN 15
2. SLN的組成 16
3. SLN的製備 17
4. SLN的輸送 19
四、研究目的 22
貳、材料與方法 23
一、藥品與化學試劑 23
二、儀器 24
三、實驗方法 25
1.定量分析 25
1.1 HPLC層析條件 25
1.2體外檢量線製作 25
1.3體外檢品檢量線之確效 25
1.4 體內檢量線製作 27
2. PK-L4溶液劑生體分佈（biodistribution）試驗 28
2.1注射劑之製備 28
2.2實驗動物 29
2.3投藥 29
2.4血漿及組織檢品之分析 29
2.5體內生體分佈（biodistribution）研究 30
3. PK-L4之SLN劑型設計 31
3.1 SLN製備方法 31
3.2奈米微粒之粒子大小、粒子分散性與表面電位之測定 35
3.3包埋率之測定 36
3.4 TEM（Transmission electron microscope）結構觀察 36
3.5 示差掃描熱分析（Differential Scanning Calorimetry, DSC） 36
4.PK-L4體外釋放試驗 37
4.1 PK-L4溶解度試驗 37
4.2 PK-L4 SLNs體外釋放試驗 37
5. PK-L4 solution與SLNs處方之細胞毒殺試驗 38
6. PK-L4 SLNs生體分佈（biodistribution）試驗 38
参、結果與討論 39
1. PK-L4分析方法建立 39
1.1 HPLC分析條件 39
1.2體外檢品檢量線 41
1.3 體外檢品檢量線之確效 42
1.4體內檢品檢量線 43
2. PK-L4 溶液劑之生體分佈（biodistribution）情形 47
2.1分室模式的分析與判定 47
2.2 PK-L4 溶液劑生體分佈（biodistribution） 48
2.3 PK-L4溶液劑之藥物動力學參數 54
3. PK-L4 SLN劑型設計 55
3.1處方之粒子大小、粒徑分散性、表面電位以及包埋率之評估 55
3.2含藥處方穿透式電子顯微鏡（TEM）結構觀察 62
3.3含藥處方的示差掃描熱分析（Differential scanning calorimetry, DSC） 64
4. PK-L4 SLN體外釋放試驗 68
5. PK-L4 solution與SLNs（without and with PEG）處方之細胞毒殺試驗 70
6. PK-L4 SLNs 生體分佈（biodistribution）試驗 72
6.1 PK-L4 SLNs生體分佈（biodistribution） 72
6.2 PK-L4 SLNs處方之藥物動力學參數 82
7. PK-L4溶液劑與SLNs生體分佈（biodistribution）及藥物動力學比較 83
肆、結論 94
伍、參考文獻 95

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